Dissociation of Infectivity from Seeding Ability in Prions with Alternate Docking Mechanism
Figure 3
Effect of accessory cofactors on the propagation of ΔPBD PrPSc molecules.
In vitro-generated ΔC-PBD, ΔN-PBD, and ΔΔ-PBD PrPSc molecules were propagated by sPMCA with autologous PrPC prepared from Chinese hamster ovary (CHO) cells. One set of reactions was supplemented with Prnp0/0 mouse brain homogenate (+cofactor), while a second set received only buffer (−cofactor). One sample of each reaction was not subjected to protease digestion (−PK), while all others were digested with 25 µg/mL proteinase K. –PK reactions for ΔC-PBD and ΔΔ-PBD were loaded with ¼ volume. PrP was detected by immunoblot (anti-PrP 27/33).